Bottom Line:
TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium.Achromobacter sp.AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.

Affiliation: Environment and Biotechnology Centre, Faculty of Life and Social Sciences, Swinburne University of Technology, PO Box 218, Melbourne, Victoria 3122, Australia. shng@swin.edu.au

ABSTRACT

Background: Achromobacter sp. AO22 (formerly Alcaligenes sp. AO22), a bacterial strain isolated from a lead-contaminated industrial site in Australia, was previously found to be resistant to moderate to high levels of mercury, copper and other heavy metals. However, the nature and location of the genetic basis for mercuric ion resistance in this strain, had not been previously identified.

Findings: Achromobacter sp. AO22 contains a functional mer operon with all four essential genes (merRTPA) and shows >99% DNA sequence identity to that of Tn501. The mer operon was present on a transposon, designated TnAO22, captured by introducing a broad-host-range IncP plasmid into Achromobacter sp. AO22 and subsequently transferring it to E. coli recipients. The transposition frequency of TnAO22 was 10-2 to 10-3 per target plasmid transferred. Analysis of TnAO22 sequence revealed it belonged to the Tn21 subgroup of the Tn3 superfamily of transposons, with the transposition module having >99% identity with Tn5051 of a Pseudomonas putida strain isolated from a water sample in New York.

Conclusion: TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium. Achromobacter sp. AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.

Mentions:
The DNA sequence identities of TnAO22 tnpA compared to its close relatives varied between 70.0% and 99.2% (Table 1). The start codon of the putative TnpA was 2 bp after termination of TnpR (data not shown), compared to 3 bp in Tn501, and it terminated within an IR. The putative TnpA is 988 amino acids long and differs from the 459 residues available for TnpA of Tn5051 at 5 positions. Alignment of the amino acid sequence of TnAO22 TnpA with 13 selected Tn3 transposases (Additional file 1 Fig. S2) and the dendrogram (Fig. 3) confirmed that TnAO22 was closest to the Tn21 subgroup of Gram negative transposons, separated from Tn3 and the cluster of transposons in Gram positive bacteria.

Mentions:
The DNA sequence identities of TnAO22 tnpA compared to its close relatives varied between 70.0% and 99.2% (Table 1). The start codon of the putative TnpA was 2 bp after termination of TnpR (data not shown), compared to 3 bp in Tn501, and it terminated within an IR. The putative TnpA is 988 amino acids long and differs from the 459 residues available for TnpA of Tn5051 at 5 positions. Alignment of the amino acid sequence of TnAO22 TnpA with 13 selected Tn3 transposases (Additional file 1 Fig. S2) and the dendrogram (Fig. 3) confirmed that TnAO22 was closest to the Tn21 subgroup of Gram negative transposons, separated from Tn3 and the cluster of transposons in Gram positive bacteria.

Bottom Line:
TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium.Achromobacter sp.AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.

Affiliation:
Environment and Biotechnology Centre, Faculty of Life and Social Sciences, Swinburne University of Technology, PO Box 218, Melbourne, Victoria 3122, Australia. shng@swin.edu.au

ABSTRACT

Background: Achromobacter sp. AO22 (formerly Alcaligenes sp. AO22), a bacterial strain isolated from a lead-contaminated industrial site in Australia, was previously found to be resistant to moderate to high levels of mercury, copper and other heavy metals. However, the nature and location of the genetic basis for mercuric ion resistance in this strain, had not been previously identified.

Findings: Achromobacter sp. AO22 contains a functional mer operon with all four essential genes (merRTPA) and shows >99% DNA sequence identity to that of Tn501. The mer operon was present on a transposon, designated TnAO22, captured by introducing a broad-host-range IncP plasmid into Achromobacter sp. AO22 and subsequently transferring it to E. coli recipients. The transposition frequency of TnAO22 was 10-2 to 10-3 per target plasmid transferred. Analysis of TnAO22 sequence revealed it belonged to the Tn21 subgroup of the Tn3 superfamily of transposons, with the transposition module having >99% identity with Tn5051 of a Pseudomonas putida strain isolated from a water sample in New York.

Conclusion: TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium. Achromobacter sp. AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.